Transfections allow for transient expression of a gene of interest in a target cell line and can be useful for short term studies of protein function. We specifically use this protocol with Lenti-X 293T cells, a cell line optimized for production of lentiviral vectors. Plaquenil toxicity macula A patient will begin taking hydroxychloroquine for rheumatoid arthritisi Chloroquine is a lysosomotropic agent that prevents endosomal acidification 1. It accumulates inside the acidic parts of the cell, including endosomes and lysosomes. This accumulation leads to inhibition of lysosomal enzymes that require an acidic pH, and prevents fusion of endosomes and lysosomes. A severe eye problem has happened with chloroquine. This may lead to lasting eyesight problems. The risk may be higher if you have some types of eye or kidney problems. The risk may also be higher with some doses of chloroquine, if you use chloroquine for longer than 5 years, or if you take certain other drugs like tamoxifen. After 2 h of pretreatment with chloroquine and 48 h of transfection with siRNA-control vector or siRNA-Atgs 3 or 7, cells were cultured in complete medium and HBSS for 6 h. RNA was then isolated by TRIzol™ Reagent Invitrogen according to the manufacturer’s protocol. Last Upload: June 10, 2016 Day 0: Seed Lenti-X 293T cells (this cell line is optimized for production of lentiviral vectors) Day 1 (pm): Transfect Cells Day 2 (am): 18h post transfection - Remove media, replace with fresh media Day 3 or more (am): Observe fluorescence, harvest cells, or perform your experiment *Pro-Tips* Different brands and lots of FBS can promote or inhibit transfection. This approach can be adapted for different cell lines and different transfection reagents. Chloroquine invitrogen Chloroquine - FDA prescribing information, side effects and uses, Chloroquine Indications, Side Effects, Warnings - Plaquenil cause low wbcCan plaquenil cause skin pigment to darkenSide effects aralen tabletsPlaquenil for asthmaRa and hydroxychloroquine Each Premo™ Autophagy Tandem Sensor RFP-GFP-LC3B Kit includes a vial of chloroquine diphosphate. Chloroquine has been demonstrated to inhibit autophagy by elevating lysosomal pH and therefore inhibiting the fusion of autophagosomes with lysosomes and preventing the subsequent lysosomal protein degradation. Premo™ Autophagy Tandem Sensor RFP-GFP-LC3B Kit. Autophagy promotes hepatocellular carcinoma cell invasion.. HCS Autophagy Assay Thermo Fisher Scientific - US. C3556 Catalase from human erythrocytes ≥90% SDS-PAGE, buffered aqueous solution, ≥30,000 units/mg protein Catalase activates the decomposition of hydrogen peroxide, a reactive oxygen species, into water and oxygen. Conclusion Together, these results suggest that Luteolin has the capacity to induce selectively apoptotic cell death both in primary cutaneous SCC cells and in metastatic SCC cells in combination with chloroquine, an inhibitor of autophagosomal degradation. Hence, Luteolin might be a promising agent for the treatment of cutaneous SCC. Chloroquine is an antimalarial drug that has been used in humans for many years. In recent years, Chloroquine has been shown to inhibit autophagy and induce apoptosis in malignant cells and thus has been tested in various experimental model systems and in human clinical trials.